Method for cryopreserving sheet-shaped cell culture

Abstract

Methods are disclosed of freezing, cryopreserving, and transferring a frozen sheet-shaped cell culture. The method of freezing includes (1) a step of immersing in a cryopreservation solution a sheet-shaped cell culture supported by a mesh-shaped support body; (2) a step of removing the cryopreservation solution adhered to the sheet-shaped cell culture, while keeping the sheet-shaped cell culture supported by the mesh-shaped support body; (3) a step of enclosing the sheet-shaped cell culture in a cold-resistant film, an upper surface and a lower surface of the sheet-shaped cell culture being covered by the mesh-shaped support body; and (4) a step of freezing the sheet-shaped cell culture.

What is claimed is: 
     
         1 . A method of freezing a sheet-shaped cell culture, comprising:
 (1) a step of immersing in a cryopreservation solution a sheet-shaped cell culture supported by a mesh-shaped support body;   (2) a step of removing the cryopreservation solution adhered to the sheet-shaped cell culture, while keeping the sheet-shaped cell culture supported by the mesh-shaped support body;   (3) a step of enclosing the sheet-shaped cell culture in a cold-resistant film, an upper surface and a lower surface of the sheet-shaped cell culture being covered by the mesh-shaped support body; and   (4) a step of freezing the sheet-shaped cell culture.   
     
     
         2 . The method according to  claim 1 , wherein in the step (1), the sheet-shaped cell culture is immersed in the cryopreservation solution for 1 to 30 minutes. 
     
     
         3 . The method according to  claim 1 , wherein in the step (2), the cryopreservation solution adhered to the sheet-shaped cell culture is removed by dropping through the mesh-shaped support body. 
     
     
         4 . The method according to  claim 1 , wherein in the step (3), the sheet-shaped cell culture is enclosed in the cold-resistant film in such a manner that a hermetically sealed state can be maintained. 
     
     
         5 . The method according to  claim 1 , wherein in the step (4), the sheet-shaped cell culture is frozen by being disposed over a liquid surface of liquid nitrogen. 
     
     
         6 . The method according to  claim 1 , wherein the step (4) is conducted after the step (3). 
     
     
         7 . A method of cryopreserving a sheet-shaped cell culture, comprising:
 (1) a step of immersing in a cryopreserving solution a sheet-shaped cell culture supported by a mesh-shaped support body;   (2) a step of removing the cryopreservation solution adhered to the sheet-shaped cell culture, while keeping the sheet-shaped cell culture supported by the mesh-shaped support body;   (3) a step of enclosing the sheet-shaped cell culture in a cold-resistant film, an upper surface and a lower surface of the sheet-shaped cell culture being covered by the mesh-shaped support body;   (4) a step of freezing the sheet-shaped cell culture; and   (5) a step of preserving the frozen sheet-shaped cell culture at a low temperature while keeping the sheet-shaped cell culture enclosed in the film.   
     
     
         8 . The method according to  claim 7 , wherein in the step (1), the sheet-shaped cell culture is immersed in the cryopreservation solution for 1 to 30 minutes. 
     
     
         9 . The method according to  claim 7 , wherein in the step (2), the cryopreservation solution adhered to the sheet-shaped cell culture is removed by dropping through the mesh-shaped support body. 
     
     
         10 . The method according to  claim 7 , wherein in the step (3), the sheet-shaped cell culture is enclosed in the cold-resistant film in such a manner that a hermetically sealed state can be maintained. 
     
     
         11 . The method according to  claim 7 , wherein in the step (4), the sheet-shaped cell culture is frozen by being disposed over a liquid surface of liquid nitrogen. 
     
     
         12 . The method according to  claim 7 , wherein the step (4) is conducted after the step (3). 
     
     
         13 . A method of transferring a sheet-shaped cell culture, comprising:
 (1) a step of immersing in a cryopreservation solution a sheet-shaped cell culture supported by a mesh-shaped support body;   (2) a step of removing the cryopreservation solution adhered to the sheet-shaped cell culture, while keeping the sheet-shaped cell culture supported by the mesh-shaped support body;   (3) a step of enclosing the sheet-shaped cell culture in a cold-resistant film, an upper surface and a lower surface of the sheet-shaped cell culture being covered by the mesh-shaped support body;   (4) a step of freezing the sheet-shaped cell culture; and   (5) a step of transferring the frozen sheet-shaped cell culture while keeping the sheet-shaped cell culture enclosed in the film.   
     
     
         14 . The method according to  claim 13 , wherein in the step (1), the sheet-shaped cell culture is immersed in the cryopreservation solution for 1 to 30 minutes. 
     
     
         15 . The method according to  claim 13 , wherein in the step (2), the cryopreservation solution adhered to the sheet-shaped cell culture is removed by dropping through the mesh-shaped support body. 
     
     
         16 . The method according to  claim 13 , wherein in the step (3), the sheet-shaped cell culture is enclosed in the cold-resistant film in such a manner that a hermetically sealed state can be maintained. 
     
     
         17 . The method according to  claim 13 , wherein in the step (4), the sheet-shaped cell culture is frozen by being disposed over a liquid surface of liquid nitrogen. 
     
     
         18 . The method according to  claim 13 , wherein the step (4) is conducted after the step (3).